Use of heteroaryl substituted N-(indole-2-carbonyl-) amides for treatment of infection

ABSTRACT

A pharmaceutical composition containing a glycogen phosphorylase inhibitor of Formula I or IA as defined herein is administered to a mammal to treat infection.

This application claims priority under 35 U.S.C. § 119(e) of U.S.application Ser. No. 60/187,605, filed Mar. 7, 2000, which applicationis hereby incorporated by reference herein.

FIELD OF THE INVENTION

This invention relates to the use of certain glycogen phosphorylaseinhibitors in the treatment of infections.

BACKGROUND OF THE INVENTION

Glycogenolysis in tissues, whereby glycogen is cleaved to releaseglucose-1-phosphate, is catalyzed by glycogen phosphorylase (GP). Inhumans, three isoforms of this enzyme have been identified: the liverisoform (HLGP), the muscle isoform (HMGP), and the brain isoform (HBGP).These isoforms are products of three separate genes and have 80-83%amino acid identity (C. B. Newgard, D. R. Littman, C. van Gendered, M.Smith, and R. J. Fletterick, J. Biol. Chem.263:3850-3857, 1988).Glycogen phosphorylase is also present in bacteria.

Glycogen phosphorylase inhibitors that have been reported to dateinclude glucose and glucose analogs (e.g., Martin, J. L. et al.,Biochemistry 1991, 30, 10101), caffeine and other purine analogs (e.g.,Kasvinsky, P. J. et al. J. Biol. Chem. 1978, 253, 3343-3351 and9102-9106), and inhibitors of the type described by Oikonomakos, N. G.et al., Protein Sci. 1999, 8, 1930-1945.

Glycogen phosphorylase inhibitors are useful in the treatment ofdiabetes mellitus. For example, International Patent publications WO96139384 and WO 96/39385, both published Dec. 12, 1996, describe use ofsubstituted N-(indole-2-carbonyl-) amides and derivatives for treatmentof diabetes. These compounds are also described as useful in treatmentof atherosclerosis, hyperinsulinemia, hypercholesterolemia,hypertension, hyperlipidemia, and in prevention of myocardial ischemicinjury.

U.S. Pat. No. 5,952,322 describes the use of glycogen phosphorylaseinhibitors, such as those described in WO 96/39384 and WO 96/39385, toreduce tissue damage associated with non-cardiac ischemia.

U.S. Pat. No. 5,882,885, issued Mar. 16, 1999 refers to antagonists andagonists of streptococcal glycogen phosphorylase as useful in thetreatment of otitis media, conjunctivitis, pneumonia, bacteremia,meningitis, sinusitis, pleural empyema and endocarditis.

SUMMARY OF THE INVENTION

The present invention relates to a method of treating or preventinginfection, e.g., bacterial, fungal, parasitic, or viral infection,comprising administering an amount of a compound of Formula I or FormulaIA that is effective in treating or preventing said infection.

Compounds of the Formula I and Formula IA have the following structures:

and the pharmaceutically acceptable salts and prodrugs thereof;

wherein:

the dotted line (---) is an optional bond;

A is —C(H)═, —C((C₁-C₄)alkyl)═ or —C(halo)═ when the dotted line (---)is a bond, or A is methylene or —CH((C₁-C₄)alkyl)— when the dotted line(---) is not a bond;

R₁, R₈ or R₉ are each independently H, halo, 4-, 6- or 7-nitro, cyano,(C₁-C₄)alkyl, (C₁-C₄)alkoxy, fluoromethyl, difluoromethyl ortrifluoromethyl;

R₂ is H;

R₃ is H or (C₁-C₅)alkyl;

R₄ is H, methyl, ethyl, n-propyl, hydroxy(C₁-C₃)alkyl,(C₁-C₃)alkoxy(C₁-C₃)alkyl, phenyl(C₁-C₄)alkyl,phenylhydroxy(C₁-C₄)alkyl, phenyl(C₁-C₄)alkoxy(C₁-C₄)alkyl, thien-2- or-3-yl(C₁-C₄)alkyl or fur-2- or -3-yl(C₁-C₄)alkyl wherein said R₄ ringsare mono-, di- or tri-substituted independently on carbon with H, halo,(C₁-C₄)alkyl, (C₁-C₄)alkoxy, trifluoromethyl, hydroxy, amino or cyano;or

R₄ is pyrid-2-, -3- or -4-yl(C₁-C₄)alkyl, thiazol-2-, -4- or-5-yl(C₁-C₄)alkyl, imidazol -1-, -2-, -4- or -5-yl(C₁-C₄)alkyl,pyrrol-2- or -3-yl(C₁-C₄)alkyl, oxazol-2-, -4- or -5-yl-(C₁-C₄)alkylpyrazol-3-, -4- or -5-yl(C₁-C₄)alkyl, isoxazol-3-, -4- or-5-yl(C₁-C₄)alkyl, isothiazol-3-, -4- or -5-yl(C₁-C₄)alkyl, pyridazin-3-or -4-yl-(C₁-C₄)alkyl, pyrimidin-2-, -4-, -5- or -6-yl(C₁-C₄)alkyl,pyrazin-2- or -3-yl(C₁-C₄)alkyl or 1,3,5-triazin-2-yl(C₁-C₄)alkyl,wherein said preceding R₄ heterocycles are optionally mono- ordi-substituted independently with halo, trifluoromethyl, (C₁-C₄)alkyl,(C₁-C₄)alkoxy, amino or hydroxy and said mono-or di-substituents arebonded to carbon;

R₅ is H, hydroxy, fluoro, (C₁-C₅)alkyl, (C₁-C₅)alkoxy, (C₁-C₆)alkanoyl,amino(C₁-C₄)alkoxy, mono-N- or di-N,N-(C₁-C₄)alkylamino(C₁-C₄)alkoxy,carboxy(C₁-C₄)alkoxy, (C₁-C₅)alkoxy-carbonyl(C₁-C₄)alkoxy,benzyloxycarbonyl(C₁-C₄)alkoxy, or carbonyloxy wherein said carbonyloxyis carbon-carbon linked with phenyl, thiazolyl, imidazolyl, 1H-indolyl,furyl, pyrrolyl, oxazolyl, pyrazolyl, isoxazolyl, isothiazolyl,pyridazinyl, pyrimidinyl, pyrazinyl or 1,3,5-triazinyl and wherein saidpreceding R₅ rings are optionally mono-substituted with halo,(C₁-C₄)alkyl, (C₁-C₄)alkoxy, hydroxy, amino or trifluoromethyl and saidmono-substituents are bonded to carbon;

R₇ is H, fluoro or (C₁-C₅)alkyl; or

R₅ and R₇ can be taken together to be oxo;

R₆ is C(O)R₁₀;

R₁₀ is piperazin-1-yl, 4-(C₁-C₄)alkylpiperazin-1-yl,4-formylpiperazin-1-yl, morpholino, thiomorpholino, 1-oxothiomorpholino,1,1-dioxo-thiomorpholino, thiazolidin-3-yl, 1 -oxo-thiazolidin-3-yl,1,1-dioxo-thiazolidin-3-yl, 2-(C₁-C₆)alkoxycarbonylpyrrolidin-1-yl,oxazolidin-3-yl or 2(R)-hydroxymethylpyrrolidin-1-yl; or

R₁₀ is 3- and/or 4-mono-or di-substituted oxazetidin-2-yl, 2-, 4-,and/or 5- mono- or di-substituted oxazolidin-3-yl, 2-, 4-, and/or 5-mono- or di- substituted thiazolidin-3-yl, 2-, 4-, and/or 5- mono- ordi- substituted 1-oxothiazolidin-3-yl, 2-, 4-, and/or 5- mono- ordi-substituted 1,1-dioxothiazolidin-3-yl, 3- and/or 4-, mono- ordi-substituted pyrrolidin-1-yl, 3-, 4-and/or 5-, mono-, di- ortri-substituted piperidin-1-yl, 3-, 4-, and/or 5- mono-, di-, ortri-substituted piperazin-1-yl, 3-substituted azetidin-1-yl, 4- and/or5-, mono- or di-substituted 1,2-oxazinan-2-yl, 3-and/or 4-mono- ordi-substituted pyrazolidin-1-yl, 4- and/or 5-, mono- or di-substitutedisoxazolidin-2-yl, 4- and/or 5-, mono- and/or di-substitutedisothiazolidin-2-yl wherein said R₁₀ substituents are independently H,halo, (C₁-C₅)-alkyl, hydroxy, amino, mono-N- ordi-N,N-(C₁-C₅)alkylamino, formyl, oxo, hydroxyimino, (C₁-C₅)alkoxy,carboxy, carbamoyl, mono-N-or di-N,N-(C₁-C₄)alkylcarbamoyl,(C₁-C₄)alkoxyimino, (C₁-C₄)alkoxymethoxy, (C₁-C₆)alkoxycarbonyl,carboxy(C₁-C₅)alkyl or hydroxy(C₁-C₅)alkyl;

R₁₂ is H, methyl, ethyl, n-propyl, hydroxy(C₁-C₃)alkyl,(C₁-C₃)alkoxy(C₁-C₃)alkyl, phenyl(C₁-C₄)alkyl,phenylhydroxy(C₁-C₄)alkyl, (phenyl)((C₁-C₄)-alkoxy)(C₁-C₄)alkyl,thien-2- or - 3-yl(C₁-C₄)alkyl or fur-2- or -3-yl(C₁-C₄)alkyl whereinsaid R₁₂ rings are mono-, di- or tri-substituted independently on carbonwith H, halo, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, trifluoromethyl, hydroxy,amino, cyano or 4,5-dihydro-1H-imidazol-2-yl; or

R₁₂ is pyrid-2-, -3- or -4-yl(C₁-C₄)alkyl, thiazol-2-, -4- or-5-yl(C₁-C₄)alkyl, imidazol-2-, -4- or - 5-yl(C₁-C₄)alkyl, pyrrol-2- or-3-yl(C₁-C₄)alkyl, oxazol-2-, -4- or -5-yl(C₁-C₄)alkyl, pyrazol-3-, -4-or -5-yl(C₁-C₄)alkyl, isoxazol-3-, -4- or -5-yl(C₁-C₄)alkyl,isothiazol-3-, 4- or -5-yl(C₁-C₄)alkyl, pyridazin-3- or-4-yl(C₁-C₄)alkyl, pyrimidin-2-, -4-, -5- or -6-yl(C₁-C₄)alkyl,pyrazin-2-or - 3-yl(C₁-C₄)alkyl, 1,3,5-triazin-2-yl(C₁-C₄)alkyl orindol-2-(C₁-C₄)alkyl, wherein said preceding R₁₂ heterocycles areoptionally mono- or di-substituted independently with halo,trifluoromethyl, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, amino, hydroxy or cyanoand said substituents are bonded to carbon; or

R₁₂ is R₁₁-carbonyloxymethyl, wherein said R₁₁ is phenyl, thiazolyl,imidazolyl, 1H-indolyl, furyl, pyrrolyl, oxazolyl, pyrazolyl,isoxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinylor 1,3,5-triazinyl and wherein said preceding R₁₁ rings are optionallymono- or di-substituted independently with halo, amino, hydroxy,(C₁-C₄)alkyl, (C₁-C₄)alkoxy or trifluoromethyl and said mono- ordi-substituents are bonded to carbon;

R₁₃ is H, methyl, ethyl, n-propyl, hydroxymethyl, or hydroxyethyl;

R₁₄ C(O)R₁₅;

R₁₅ is morpholino, thiomorpholino, 1-oxothiomorpholino,1,1-dioxothiomorpholino, thiazolidin-3-yl, 1-oxothiazolidin-3-yl,1,1-dioxothiazolidin-3-yl, pyrrolidin-1-yl, piperidin-1-yl,piperazin-1-yl, piperazin-4-yl, azetidin-1-yl, 1,2-oxazinan-2-yl,pyrazolidin-1-yl, isoxazolidin-2-yl, isothiazolidin-2-yl,1,2-oxazetidin-2-yl, oxazolidin-3-yl, 3,4-dihydroisoquinolin-2-yl,1,3-dihydroisoindol-2-yl, 3,4-dihydro-2H-quinol-1 -yl,2,3-dihydro-benzo[1,4]oxazin4-yl, 2,3-dihydro-benzo[1,4]-thiazine-4-yl,3,4-dihydro-2H-quinoxalin-1 -yl, 3,4-dihydrobenzo[c][1,2]oxazin-1-yl,1,4-dihydro-benzo[d][1,2]oxazin-3-yl,3,4-dihydro-benzo[e][1,2]-oxazin-2-yl, 3H-benzo[d]isoxazol-2-yl,3H-benzo[c]isoxazol-1-yl or azepan-1-yl,

wherein said R₁₅ rings are optionally mono-, di- or tri-substitutedindependently with halo, (C₁-C₅)alkyl, (C₁-C₅)alkoxy, hydroxy, amino,mono-N- or di-N,N-(C₁-C₅)alkylamino, formyl, carboxy, carbamoyl, mono-N-or di-N,N-(C₁-C₅)alkylcarbamoyl, (C₁-C₆)alkoxy(C₁-C₃)alkoxy,(C₁-C₅)alkoxycarbonyl, benzyloxycarbonyl,(C₁-C₅)alkoxycarbonyl(C₁-C₅)alkyl, (C₁-C₄)alkoxycarbonylamino,carboxy(C₁-C₅)alkyl, carbamoyl(C₁-C₅)alkyl, mono-N- ordi-N,N-(C₁-C₅)alkylcarbamoyl(C₁-C₅)alkyl, hydroxy(C₁-C5)alkyl,(C₁-C₄)alkoxy(C₁-C₄)alkyl, amino(C₁-C₄)alkyl, mono-N- ordi-N,N-(C₁-C₄)alkylamino(C₁-C₄)alkyl, oxo, hydroxyimino or(C₁-C₆)alkoxyimino and wherein no more than two substituents areselected from oxo, hydroxyimino or (C₁-C₆)alkoxyimino and oxo,hydroxyimino or (C₁-C₆)alkoxyimino are on nonaromatic carbon; and

wherein said R₁₅ rings are optionally additionally mono- ordi-substituted independently with (C₁-C₅)alkyl or halo.

A group of preferred compounds of Formula I consists of those compoundswherein:

R₁ is 5-H, 5-halo, 5-methyl or 5-cyano;

R₈ and R₉ are each independently H or halo;

A is —C(H)═;

R₂ and R₃ are H;

R₄ is phenyl(C₁-C₂)alkyl wherein said phenyl groups are mono-, di- ortri-substituted independently with H or halo or mono- or di- substitutedindependently with H, halo, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,trifluoromethyl, hydroxy, amino or cyano; or

R₄ is thien-2- or -3-yl(C₁-C₂)alkyl, pyrid-2-, -3- or -4-yl(C₁-C₂)alkyl,thiazol-2-, -4- or -5-yl(C₁-C₂)alkyl, imidazol -1-, -2-, -4- or-5-yl(C₁-C₂)alkyl, fur-2- or -3-yl(C₁-C₂)alkyl, pyrrol-2- or-3-yl(C₁-C₂)alkyl, oxazol-2-, 4- or -5-yl-(C₁-C₂)alkyl, pyrazol-3-, -4-or -5-yl(C₁-C₂)alkyl, isoxazol-3-, 4- or -5-yl(C₁-C₂)alkyl wherein saidpreceding R₄ heterocycles are optionally mono- or di-substitutedindependently with halo, trifluoromethyl, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,amino or hydroxy and said mono- or di-substituents are bonded to carbon;

R₅ is hydroxy; and

R₇is H.

Within the above group of preferred compounds of Formula I is a secondgroup of especially preferred compounds wherein

the carbon atom labelled a has (S) stereochemistry;

the carbon atom labelled b has (R) stereochemistry;

R₄ is phenyl(C₁-C₂)alkyl, thien-2-yl-(C₁-C₂)alkyl,thien-3-yl-(C₁-C₂)alkyl, fur-2-yl-(C₁-C₂)alkyl or fur-3-yl-(C₁-C₂)alkylwherein said rings are mono- or di- substituted independently with H orfluoro; and

R₁₀ is morpholino, 4-(C₁-C₄)alkylpiperazin-1-yl, 3-substitutedazetidin-1-yl, 3- and/or 4-, mono- or di-substituted pyrrolidin-1-yl, 4-and/or 5- mono- or di-substituted isoxazolidin-2-yl, 4- and/or 5-, mono-or di-substituted 1,2-oxazinan-2-yl wherein said substituents are eachindependently H, halo, hydroxy, amino, mono-N- ordi-N,N-(C₁-C₆)alkylamino, oxo, hydroxyimino or alkoxy.

Within the above group of especially preferred compounds are theparticularly preferred compounds:

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-(2R)-hydroxy-3-(4-methylpiperazin-1-yl)-3-oxo-propyl]-amidehydrochloride,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-(2R)-hydroxy-3-(3-hydroxyazetidin-1-yl)-3-oxo-propyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid((1S)-benzyl-(2R)-hydroxy-3-isoxazolidin-2-yl-3-oxo-propyl)-amide,

5-Chloro-1H-indole-2-carboxylic acid((1S)-benzyl-(2R)-hydroxy-3-[1,2]oxazinan-2-yl-3-oxo-propyl)-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-(2R)-hydroxy-3-((3S)-hydroxypyrrolidin-1-yl)-3-oxo-propyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-3-((3S,4S)-dihydroxy-pyrrolidin-1-yl)-(2R)-hydroxy-3-oxo-propyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-3-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-(2R)-hydroxy-3-oxo-propyl]-amide;and

5-Chloro-1H-indole-2-carboxylic acid((1S)-benzyl-(2R)-hydroxy-3-morpholin-4-yl-3-oxo-propyl)-amide.

Within the above group of especially preferred compounds of Formula Iare compounds wherein:

a.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₄ is benzyl; and

R₁₀ is 4-methylpiperazin-1-yl;

b.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₄ is benzyl; and

R₁₀ is 3-hydroxyazetidin-1-yl;

c.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₄ is benzyl; and

R₁₀ is isoxazolidin-2-yl;

d.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₄ is benzyl; and

R₁₀ is (1,2)-oxazinan-2-yl;

e.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₄ is benzyl; and

R₁₀ is 3(S)-hydroxypyrrolidin-1-yl;

f.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₄ is benzyl; and

R₁₀ is (3S,4S)-dihydroxypyrrolidin-1-yl;

g.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₄ is benzyl; and

R₁₀ is cis-3,4-dihydroxypyrrolidin-1-yl; and

h.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₄ is benzyl; and

R₁₀ is morpholino.

Another group of preferred compounds of Formula I are those wherein:

R₁ is H, halo, methyl or cyano;

R₈ and R₉ are each independently H or halo;

A is —C(H)═;

R₂ and R₃ are H;

R₄ is phenyl(C₁-C₂)alkyl wherein said phenyl groups are mono-, di- ortri-substituted independently with H or halo or mono- or di- substitutedindependently with H, halo, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,trifluoromethyl, hydroxy, amino or cyano; or

R₄ is thien-2- or -3-yl(C₁-C₂)alkyl, pyrid-2-, -3- or -4-yl(C₁-C₂)alkyl,thiazol-2-, -4- or -5-yl(C₁-C₂)alkyl, imidazol -1-, -2-, -4- or-5-yl(C₁-C₂)aikyl, fur-2- or -3-yl(C₁-C₂)alkyl, pyrrol-2- or-3-yl(C₁-C₂)alkyl, oxazol-2-, -4- or -5-yl-(C₁-C₂)alkyl, pyrazol-3-, -4-or -5-yl(C₁-C₂)alkyl, isoxazol-3-, -4- or -5-yl(C₁-C₂)alkyl wherein saidpreceding R₄ heterocycles are optionally mono- or di-substitutedindependently with halo, trifluoromethyl, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,amino or hydroxy and said mono- or di-substituents are bonded to carbon;

R₅ is fluoro, (C₁-C₄)alkyl, (C₁-C₅)alkoxy, amino(C₁-C₄)alkoxy, mono-N-or di-N,N-(C₁-C₄)alkylamino(C₁-C₄)alkoxy, carboxy(C₁-C₄)aIkoxy,(C₁-C₅)alkoxy-carbonyl(C₁-C₄)alkoxy, benzyloxycarbonyl(C₁-C₄)alkoxy; and

R₇ is H, fluoro or (C₁-C₆)alkyl.

A group of preferred compounds of Formula IA consists of those compoundswherein:

R₁ is 5-H, 5-halo, 5-methyl, 5-cyano or 5-trifluoromethyl;

R₈ and R₉ are each independently H or halo;

A is —C(H)═;

R₂ and R₃ are H;

R₁₂ is H, methyl, phenyl(C₁-C₂)alkyl, wherein said phenyl groups aremono- or di-substituted independently with H, halo, (C₁-C₄)alkyl,(C₁-C₄)alkoxy, trifluoromethyl, hydroxy, amino or cyano and wherein saidR₁₂ groups are optionally additionally mono-substituted with halo; or

R₁₂ is thien-2- or -3-yl(C₁-C₂)alkyl, pyrid-2-, -3- or-4-yl(C₁-C₂)alkyl, thiazol-2-, -4- or -5-yl(C₁-C₂)alkyl, imidazol-2-,-4- or -5-yl(C₁-C₂)alkyl, fur-2- or -3-yl(C₁-C₂)alkyl, pyrrol-2- or-3-yl(C₁-C₂)alkyl, oxazol-2-, -4- or -5-yl(C₁-C₂)alkyl, pyrazol-3-, -4-or -5-yl(C₁-C₂)alkyl, isoxazol-3-, -4- or -5-yl(C₁-C₂)alkyl,isothiazol-3-, -4- or -5-yl(C₁-C₂)alkyl, pyridazin-3- or-4-yl(C₁-C₂)alkyl, pyrimidin-2-, -4-, -5- or -6-yl(C₁-C₂)alkyl,pyrazin-2- or -3-yl(C₁-C₂)alkyl or 1,3,5-triazin-2-yl(C₁-C₂)alkylwherein said preceding R₁₂ heterocycles are optionally mono- ordi-substituted independently with halo, trifluoromethyl, (C₁-C₄)alkyl,(C₁-C₄)alkoxy, amino or hydroxy and said mono- or di-substituents arebonded to carbon; and

R₁₃ is H.

Within the above group of preferred compounds of Formula IA is a groupof especially preferred compounds wherein:

R₁₂ is H, phenyl(C₁-C₂)alkyl, thien-2- or -3-yl(C₁-C₂)alkyl, fur-2- or-3-yl(C₁-C₂)alkyl wherein said R₁₂ rings are mono- or di-substitutedindependently with H or fluoro; and

R₁₅ is morpholino, thiomorpholino, 1-oxothiomorpholino,1,1-dioxothiomorpholino, thiazolidin-3-yl, 1-oxothiazolidin-3-yl,1,1-dioxothiazolidin-3-yl, pyrrolidin-1-yl, piperidin-1-yl,piperazin-1-yl, piperazin-4-yl, azetidin-1-yl, 1,2-oxazinan-2-yl,isoxazolidin-2-yl, isothiazolidin-2-yl, 1,2-oxazetidin-2-yl,oxazolidin-3-yl, 1,3-dihydroisoindol-2-yl, or azepan-1-yl,

wherein said R₁₅ rings are optionally mono- or di-substitutedindependently with halo, (C₁-C₅)alkyl, (C₁-C₅)alkoxy, hydroxy, amino,mono-N-or di-N,N-(C₁-C₅)alkylamino, formyl, carboxy, carbamoyl, mono-N-or di-N,N-(C₁-C₅)alkylcarbamoyl, (C₁-C₅)alkoxycarbonyl,hydroxy(C₁-C₅)alkyl, amino(C₁-C₄)alkyl, mono-N- ordi-N,N-(C₁-C₄)alkylamino(C₁-C₄)alkyl, oxo, hydroxyimino or(C₁-C₆)alkoxyimino with the proviso that only the R₁₅ heterocyclesthiazolidin-3-yl, pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl,piperazin-4-yl, azetidin-1-yl, 1,2-oxazinan-2-yl, isoxazolidin-2-yl, oroxazolidin-3-yl are optionally mono- or di-substituted with oxo,hydroxyimino, or (C₁-C₆)alkoxyimino; and

wherein said R₁₅ rings are optionally additionally mono- ordi-substituted independently with (C₁-C₅)alkyl.

Within the above group of especially preferred compounds are thecompounds:

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-(3-hydroxyimino-pyrrolidin-1 -yl)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[2-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[2-((3S,4S)-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid [2-(1,1-dioxc-thiazolidin-3-yl)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid(2-oxo-2-thiazolidin-3-yl-ethyl)-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-(4-fluoro-benzyl)-2-(4-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-((3RS)-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[2-oxo-2-((1RS)-oxo-1-thiazolidin-3-yl)-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-(2-fluoro-benzyl)-2-(4-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-((3S,4S)-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-(3-hydroxy-azetidin-1 -yi)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-(3-hydroxyimino-azetidin-1 -yl)-2-oxo-ethyl]-amide,

5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-(4-hydroxyimino-piperidin-1 -yl)-2-oxo-ethyl]-amide, and

5-Chloro-1H-indole-2-carboxylic acid [1-benzyl-2-(3-hydroxypyrrolidin-1-yl)-2-oxo-ethyl]amide.

Within the above group of especially preferred compounds of Formula IAis a group of particularly preferred compounds wherein:

R₁₂ is H; and

R₁₅ is thiazolidin-3-yl, 1-oxo-thiazolidin-3-yl,1,1-dioxo-thiazolidin-3-yl or oxazolidin-3-yl or said R₁₅ substituentsoptionally mono- or di-substituted independently with carboxy,(C₁-C₅)alkoxycarbonyl, hydroxy(C₁-C₃)alkyl, amino(C₁-C₃)alkyl, mono-N-or di-N,N-(C₁-C₃)alkylamino(C₁-C₃)alkyl or

R₁₅ is mono- or di-substituted pyrrolidin-1-yl wherein said substituentsare independently carboxy, (C₁-C₅)alkoxycarbonyl, (C₁-C₅)alkoxy,hydroxy, hydroxy(C₁-C₃)alkyl, amino, amino(C₁-C₃)alkyl, mono-N- ordi-N,N-(C₁-C₃)alkylamino(C₁-C₃)alkyl or mono-N- ordi-N,N-(C₁-C₄)alkylamino; and

the R₁₅ rings are optionally additionally independently disubstitutedwith (C₁-C₅)alkyl.

Preferred compounds within the immediately preceding group of compoundsare those wherein:

a.

R₁ is 5-chloro;

R₈ and R₉ are H; and

R₁₅ is cis-3,4-dihydroxy-pyrrolidin-1-yl;

b.

R₁ is 5-chloro;

R₈ and R₉ are H; and

R₁₅ is (3S,4S)-dihydroxy-pyrrolidin-1-yl;

c.

R₁ is 5-chloro;

R₈ and R₉ are H; and

R₁₅ is 1,1 -dioxo-thiazolidin-3-yl;

d.

R₁ is 5-chloro;

R₈ and R₉ are H; and

R₁₅ is thiazolidin-3-yl; and

e.

R₁ is 5-chloro;

R₈ and R₉ are H; and

R₁₅ is 1-oxo-thiazolidin-3-yl.

Within the above group of especially preferred compounds of Formula IAis another group of particularly preferred compounds wherein:

R₁₅ is phenylmethyl, thien-2- or -3-ylmethyl wherein said R₁₅ rings areoptionally mono- or di-substituted with fluoro; and

R₁₅ is thiazolidin-3-yl, 1-oxo-thiazolidin-3-yl,1,1-dioxo-thiazolidin-3-yl or oxazolidin-3-yl or said R₁₅ substituentsoptionally mono- or di-substituted independently with carboxy or(C₁-C₅)alkoxycarbonyl, hydroxy(C₁-C₃)alkyl, amino(C₁-C₃)alkyl or mono-N-or di-N,N-(C₁-C₃)alkylamino(C₁-C₃)alkyl

or R₁₅ is mono- or di-substituted azetidin-1-yl or mono- ordi-substituted pyrrolidin-1-yl or mono- or di-substituted piperidin-1-ylwherein said substituents are independently carboxy,(C₁-C₅)alkoxycarbonyl, hydroxy(C₁-C₃)alkyl, amino(C₁-C₃)alkyl, mono-N-or di-N,N-(C₁-C₃)alkylamino(C₁-C₃)alkyl, hydroxy, (C₁-C₅)alkoxy, amino,mono-N- or di-N,N-(C₁-C₅)alkylamino, oxo, hydroxyimino or(C₁-C₅)alkoxyimino; and

the R₁₅ rings are optionally additionally mono- or di-substitutedindependently with (C₁-C₅)alkyl.

Preferred compounds within the immediately preceding group ofparticularly preferred compounds of Formula IA are compounds wherein

a.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₁₂ is 4-fluorobenzyl;

R₁₅ is 4-hydroxypiperidin-1-yl; and

the stereochemistry of carbon (a) is (S);

b.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₁₂ is benzyl;

R₁₅ is 3-hydroxypiperidin-1-yl; and

the stereochemistry of carbon (a) is (S);

c.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₁₂ is benzyl;

R₁₅ is cis-3,4-dihydroxy-pyrrolidin-1-yl; and

the stereochemistry of carbon (a) is S;

d.

R₁ is 5-chloro;

R₈ and R₉ are H; R₁₂ is benzyl;

R₁₅ is 3-hydroxyimino-pyrrolidin-1-yl; and

the stereochemistry of carbon (a) is (S);

e.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₁₂ is 2-fluorobenzyl;

R₁₅ is 4-hydroxypiperidin-1-yl; and

the stereochemistry of carbon (a) is (S);

f.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₁₂ is benzyl;

R₁₅ is (3S,4S)-dihydroxy-pyrrolidin-1-yl; and

the stereochemistry of carbon (a) is (S);

g.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₁₂ is benzyl;

R₁₅ is 3-hydroxy-azetidin-1-yl; and

the stereochemistry of carbon (a) is (S);

h.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₁₂ is benzyl;

R₁₅ is 3-hydroxyimino-azetidin-1-yl; and

the stereochemistry of carbon (a) is (S); and

i.

R₁ is 5-chloro;

R₈ and R₉ are H;

R₁₂ is benzyl;

R₁₅ is 4-hydroxyimino-piperidin-1-yl; and

the stereochemistry of carbon (a) is (S).

The glycogen phosphorylase inhibitor of formula I or IA is employed totreat bacterial infections and protozoa infections and disorders relatedto such infections that include the following: pneumonia, otitis media,sinusitus, bronchitis, tonsillitis, and mastoiditis related to infectionby Streptococcus pneumoniae, Haemophilus influenzae, Moraxellacatarrhalis, Staphylococcus aureus, or Peptostreptococcus spp.;pharynigitis, rheumatic fever, and glomerulonephritis related toinfection by Streptococcus pyogenes, Groups C and G streptococci,Clostridium diptheriae, or Actinobacillus haemolyticum; respiratorytract infections related to infection by Mycoplasma pneumoniae,Legionella pneumophila, Streptococcus pneumoniae, Haemophilusinfluenzae, or Chlamydia pneumoniae; uncomplicated skin and soft tissueinfections, abscesses and osteomyelitis, and puerperal fever related toinfection by Staphylococcus aureus, coagulase-positive staphylococci(i.e., S. epidermidis, S. hemolyticus, etc.), Streptococcus pyogenes ,Streptococcus agalactiae, Streptococcal groups C-F (minute-colonystreptococci), viridans streptococci, Corynebacterium minutissimum,Clostridium spp., or Bartonella henselae; uncomplicated acute urinarytract infections related to infection by Staphylococcus saprophyticus orEnterococcus spp.; urethritis and cervicitis; and sexually transmitteddiseases related to infection by Chlamydia trachomatis, Haemophilusducreyi, Treponema pallidum, Ureaplasma urealyticum, or Neiserriagonorrheae; toxin diseases related to infection by S. aureus (foodpoisoning and Toxic shock syndrome), or Groups A, B, and C streptococci;ulcers related to infection by Helicobacter pylori; systemic febrilesyndromes related to infection by Borrelia recurrentis; Lyme diseaserelated to infection by Borrelia burgdorferi; conjunctivitis, keratitis,and dacrocystitis related to infection by Chlamydia trachomatis,Neisseria gonorrhoeae, S. aureus, S. pneumoniae, S. pyogenes, or H.influenzae; disseminated Mycobacterium avium complex (MAC) diseaserelated to infection by Mycobacterium avium, or Mycobacteriumintracellulare; gastroenteritis related to infection by Campylobacterjejuni; intestinal protozoa related to infection by Cryptosporidiumspp.; odontogenic infection related to infection by viridansstreptococci; persistent cough related to infection by Bordetellapertussis; gas gangrene related to infection by Clostridium perfringensor Bacteroides spp.; atherosclerosis related to infection byHelicobacter pylori, Chlamydia pneumoniae, or Mycoplasma pneumoniae,dysentery related to infection by Shigella dysenteriae, and symptoms ofinfection by enterotoxigenic E. coli or Mycobacterium tuberculosis.Bacterial infections and protozoa infections and disorders related tosuch infections that may be treated or prevented in animals include thefollowing: bovine respiratory disease related to infection byPasteurella haemolyticus, P. multocida, Mycoplasma bovis, or Bordetellaspp.; cow enteric disease related to infection by E. coli or protozoa(i.e., coccidia, cryptosporidia, etc.); dairy cow mastitis related toinfection by Staph. aureus, Strep. uberis, Strep. agalactiae, Strep.dysgalactiae, Klebsiella spp., Corynebacterium, or Enterococcus spp.;swine respiratory disease related to infection by Actinobacifluspleuropneumoniae, P. multocida, or Mycoplasma spp.; swine entericdisease related to infection by E. coli, Lawsonia intracellularis,Salmonella, or Serpulina hyodysenteriae; cow footrot related toinfection by Fusobacterium spp.; cow metritis related to infection by E.coli; cow hairy warts related to infection by Fusobacterium necrophorumor Bacteroides nodosus; cow pink-eye related to infection by Moraxellabovis; cow premature abortion related to infection by protozoa (i.e.neosporium); urinary tract infection in dogs and cats related toinfection by E. coli; skin and soft tissue infections in dogs and catsrelated to infection by Staph. epidermidis, Staph intermedius, coagulaseneg. Staph. or P. multocida; and dental or mouth infections in dogs andcats related to infection by Alcaligenes spp., Bacteroides spp.,Clostridium spp., Enterobacter spp., Eubacterium, Peptostreptococcus,Porphyromonas, or Prevotella. The invention also encompasses treatmentof bacteremia, meningitis, pleural empyema, malaria, river blindness,toxoplasmosis, and endocarditis. Other bacterial infections and protozoainfections and disorders related to such infections that may be treatedor prevented in accord with the method of the present invention arereferred to in J. P. Sanford et al., “The Sanford Guide To AntimicrobialTherapy,” 26th Edition, (Antimicrobial Therapy, Inc., 1996).

In one embodiment, the infection that is treated according to theinvention is mediated by an organism that requires glycogen, or glucosethat results from the breakdown of glycogen, as a source of energyand/or carbon supply.

In another embodiment, the glycogen phosphorylase inhibitor isadministered in an amount that reduces or eliminates infectionsufficiently to reduce complications, including long-term complications,that can be associated with the infection. These complications include,but are not limited to asthma, and cerebrovascular disease.

In an alternative embodiment, the present invention relates to apharmaceutical composition for the treatment of bacterial infectioncomprising an amount of a compound of Formula I or IA effective to treatsaid infection in combination with a pharmaceutically acceptablecarrier.

In another embodiment, a glycogen phosphorylase inhibitor isadministered to treat Chiamydia pneumoniae infection.

DETAILED DESCRIPTION OF THE INVENTION

All patents, patent publications, and literature references cited hereinare hereby incorporated by reference.

It is intended that reference to particular compounds herein beinterpreted to mean that the pharmaceutically acceptable anionic orcationic salts and prodrugs of those compounds may also be employed.

Methods for making the glycogen phosphorylase inhibitors describedherein are described in detail in U.S. Pat. No. 5,952,322 and in WO96/39384 and WO 96/39385.

The term glycogen phosphorylase inhibitor refers to a substance or agentor combination of substances and/or agents which reduces, retards, oreliminates the enzymatic action of glycogen phosphorylase. The currentlyknown enzymatic action of glycogen phosphorylase is the degradation ofglycogen by catalysis of the reversible reaction of a glycogenmacromolecule and inorganic phosphate to glucose-1-phosphate and aglycogen macromolecule which is one glucosyl residue shorter than theoriginal glycogen macromolecule (forward direction of glycogenolysis).

The term “treating” as used herein includes preventative (e.g.,prophylactic) and palliative treatment.

By halo is meant chloro, bromo, iodo, or fluoro.

By alkyl is meant straight chain or branched saturated hydrocarbon.Exemplary of such alkyl groups (assuming the designated lengthencompasses the particular example) are methyl, ethyl, propyl,isopropyl, butyl, sec-butyl, tertiary butyl, pentyl, isopentyl, hexyland isohexyl.

By alkoxy is meant straight chain or branched saturated alkyl bondedthrough an oxy. Exemplary of such alkoxy groups (assuming the designatedlength encompasses the particular example) are methoxy, ethoxy, propoxy,isopropoxy, butoxy, isobutoxy, tertiary butoxy, pentoxy, isopentoxy,hexoxy and isohexoxy.

The expression “pharmaceutically-acceptable anionic salt” refers tonontoxic anionic salts containing anions such as (but not limited to)chloride, bromide, iodide, sulfate, bisulfate, phosphate, acetate,maleate, fumarate, oxalate, lactate, tartrate, citrate, gluconate,methanesulfonate and 4-toluene-sulfonate.

The expression “pharmaceutically-acceptable cationic salt” refers tonontoxic cationic salts such as (but not limited to) sodium, potassium,calcium, magnesium, ammonium or protonated benzathine(N,N′-dibenzylethylenediamine), choline, ethanolamine, diethanolamine,ethylenediamine, meglamine (N-methyl-glucamine), benethamine(N-benzylphenethylamine), piperazine or tromethamine(2-amino-2-hydroxymethyl-1,3-propanediol).

The expression “prodrug” refers to compounds that are drug precursors,which following administration, release the drug in vivo via somechemical or physiological process (e.g., a prodrug on being brought tothe physiological pH is converted to the desired drug form). Exemplaryprodrugs upon cleavage release the corresponding free acid, and suchhydrolyzable ester-forming residues of the compounds of Formula I and IAinclude but are not limited to carboxylic acid substituents (e.g., R₁₀contains carboxy) wherein the free hydrogen is replaced by (C₁-C₄)alkyl,(C₂-C₁₂)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having from 4 to 9carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having from 5 to 10 carbonatoms, alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms,1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms,1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms,N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms,1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms,3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl,di-N,N-(C₁-C₂)alkylamino(C₂-C₃)alkyl (such as α-dimethylaminoethyl),carbamoyl-(C₁-C₂)alkyl, N,N-di(C₁-C₂)alkylcarbamoyl-(C₁-C₂)alkyl andpiperidino-, pyrrolidino- or morpholino(C₂-C₃)alkyl.

Other exemplary prodrugs release an alcohol of Formula I of IA whereinthe free hydrogen of the hydroxy substituent (e.g., R₅ is hydroxy) isreplaced by (C₁-C₆)alkanoyloxymethyl, 1-((C₁-C₆)alkanoyloxy)ethyl,1-methyl-1-((C₁-C₆)alkanoyloxy)ethyl, (C₁-C₆)alkoxycarbonyloxymethyl,N-(C₁-C₆)alkoxycarbonylaminomethyl, succinoyl, (C₁-C₆)alkanoyl,α-amino(C₁-C₄)alkanoyl, arylactyl and α-aminoacyl, orα-aminoacyl-α-aminoacyl wherein said α-aminoacyl moieties areindependently any of the naturally occurring L-amino acids found inproteins, P(O)(OH)₂, —P(O)(O(C₁-C₆)alkyl)₂ or glycosyl (the radicalresulting from detachment of the hydroxyl of the hemiacetal of acarbohydrate).

Other exemplary prodrugs include but are not limited to derivatives ofFormula I or IA wherein R₂ is a free hydrogen which is replaced byR-carbonyl, RO-carbonyl, NRR′-carbonyl where R and R′ are eachindependently ((C₁-C₁₀)alkyl, (C₃-C₇)cycloalkyl, benzyl, or R-carbonylis a natural α-aminoacyl or natural α-aminoacyl-natural α-aminoacyl,—C(OH)C(O)OY wherein Y is H, (C₁-C₆)alkyl or benzyl, —C(OY₀)Y₁ whereinY₀ is (C₁-C₄) alkyl and Y₁ is ((C₁-C₆)alkyl, carboxy(C₁-C₆)alkyl,amino(C₁-C₄)alkyl or mono-N- or di-N,N-(C₁-C₆)alkylaminoalkyl, —C(Y₂)Y₃wherein Y₂ is H or methyl and Y₃ is mono-N— or di-N,N-(C₁-C₆)alkylamino,morpholino, piperidin-1-yl or pyrrolidin-1-yl.

Other exemplary prodrugs include but are not limited to derivatives ofFormula I or IA bearing a hydrolyzable moiety at R₃, which release acompound of formula I or IA wherein R₃ is a free hydrogen on hydrolysis.Such hydrolyzable moieties at R₃ are/include 1-hydroxy(C₁-C₆)alkyl or1-hydroxy-1-phenylmethyl.

Other exemplary prodrugs include cyclic structures such as compounds ofFormula I or IA wherein R₂ and R₃ are a common carbon, thus forming afive-membered ring. The linking carbon may be mono- or di-substitutedindependently with H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl or phenyl.Alternatively, R₃ and R₅ may be taken together to form an oxazolidinering and the number 2 carbon of the oxazolidine ring may be mono- ordi-substituted independently with H, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl orphenyl.

Mammals treated according to the invention include but are not limitedto humans. In one embodiment, the mammal is a companion animal, such asa dog or cat.

The chemist of ordinary skill will recognize that certain compounds ofFormula I and IA contain one or more atoms which may be in a particularstereochemical or geometric configuration, giving rise to stereoisomersand configurational isomers. Examples of such atoms are the carbon atomslabelled (a) and (b) in Formula I, and the carbon atom labelled (a) inFormula 1A. All such isomers and mixtures thereof are included in themethod and composition of the invention. Hydrates of the compounds ofFormula I and IA are also included.

The compounds of Formula I and IA have asymmetric carbon atoms andtherefore are enantiomers or diastereomers. Diasteromeric mixtures canbe separated into their individual diastereomers on the basis of theirphysical chemical differences by methods known per se., for example, bychromatography and/or fractional crystallization. Enantiomers can beseparated by converting the enantiomeric mixture into a diasteromericmixture by reaction with an appropriate optically active compound (e.g.,alcohol), separating the diastereomers and converting (e.g.,hydrolyzing) the individual diastereomers to the corresponding pureenantiomers. All such isomers, including diastereomers, enantiomers andmixtures thereof are considered as part of the method and composition ofthis invention. Use of any tautomers of compounds of Formula I and IA isalso encompassed by the invention.

Although many compounds employed in this invention are not ionizable atphysiological conditions, some of the compounds employed in thisinvention are ionizable at physiological conditions. Thus, for examplesome of the compounds employed in this invention are acidic and theyform a salt with a pharmaceutically acceptable cation. All such saltsare within the scope of the method and composition of this invention andthey can be prepared by conventional methods. For example, they can beprepared simply by contacting the acidic and basic entities, usually ina stoichiometric ratio, in either an aqueous, non-aqueous or partiallyaqueous medium, as appropriate. The salts are recovered either byfiltration, by precipitation with a non-solvent followed by filtration,by evaporation of the solvent, or, in the case of aqueous solutions, bylyophilization, as appropriate.

In addition, some of the compounds employed in this invention are basic,and they form a salt with a pharmaceutically acceptable anion. All suchsalts are within the scope of the method and composition of thisinvention and they can be prepared by conventional methods. For example,they can be prepared simply by contacting the acidic and basic entities,usually in a stoichiometric ratio, in either an aqueous, non-aqueous orpartially aqueous medium, as appropriate. The salts are recovered eitherby filtration, by precipitation with a non-solvent followed byfiltration, by evaporation of the solvent, or, in the case of aqueoussolutions, by lyophilization, as appropriate.

In addition, use of any hydrates or solvates of compounds of formula Ior IA is also within the scope of the invention.

Glycogen phosphorylase inhibition is readily determined by those skilledin the art according to standard assays.

Methods for obtaining glycogen phosphorylases, and assays fordetermining glycogen phosphorylase inhibition are described below. Othersources of glycogen phosphorylase, and other glycogen phosphorylaseinhibition assays, are known in the art. For example the glycogenphosphorylase of U.S. Pat. No. 5,882,885 may also be employed.

Purification, Expression, and Assaying of Glycogen Phosphorylase fromPathogens:

Methods and strategies for cloning and expressing glycogen phosphorylasefrom bacteria or other pathogens are known in the art of molecularbiology. In general, primers are designed to encompass the desiredglycogen phosphorylase. The specific PCR product containing the desiredglycogen phosphorylase is amplified, purified, and inserted into anappropriate plasmid to allow expression of the heterologous protein inE. coli under the control of a regulated promoter (e.g., trp or lac). Tosimplify purification, a host cell is preferably employed that lacksphoA, an endogenous phosphatase that is known to interfere with theassaying of glycogen phosphorylase. Purification of the enzyme can beaccomplished by the procedure of Seok, et. al. (Seok, et al., 1997, J.Biol. Chem. 272:26511-26521) or by using tags (e.g., his tags or proteinfusions) that aid in purification. Assay of glycogen phosphorylases fromdifferent bacteria may require optimization of the reaction conditionsfollowing purification of the enzyme activity. The assay can be run ineither a forward or reverse manner (the forward direction monitorsproduction of glucose-i-phosphate from glycogen or another substrate;the reverse reaction measures production of glycogen fromglucose-1-phosphate by monitoring the release of inorganic phosphate).

To assess the activity of a compound for general antibacterial activity,those skilled in the art can follow guidelines developed by the NationalCommittee for Clinical Laboratory Standards (Methods for dilutionantimicrobial susceptibility tests for bacteria that growaerobically—4^(th) Edition; Approved Standard. NCCLS document M7-A4(ISBN 1-56238-309-4) 1997; Methods for antimicrobial susceptibilitytesting of anaerobic bacteria—3^(rd) Edition; Approved Standards. NCCLSdocument M11-A3 (ISBN 1-56238-210-1) 1993). Assays for determiningantibacterial activity against intracellular pathogens vary according tothe proscribed literature for each organism. Some specific examples anddetails are described below. Tests for determining activity againstother organisms are known in the art.

Methodology for Testing of Mycobacterium avium:

Both agar and broth dilution assays can be performed to determine the invitro susceptibility (MIC) of Mycobacterium avium complex (Inderlied, C.B. et al., Antimicrob. Agents Chemother., 1987, 31:1697-1702.). Fordetermining the susceptibility of M. avium while growing intracellularlyin human monocytes, 100 μL of a well-dispersed suspension of M. aviumcells (final concentration of ˜5×10⁷ cells/mL) is added to each well ofa 24-well tissue culture plate containing monocytes (as described byBermudez, L. E., et al., Antimicrob Agents Chemother., 1996,40:546-551). After 4 hours, quantitative plate counts of lysedmacrophage monolayers are performed to establish the baseline of M.avium cells/mL within the macrophages. Infected monclayers are thentreated with compound at different concentrations; compound and mediumare replenished daily for 4 days. After the 4-day treatment period, themedium is removed and the monolayers are lysed using ice-cold sterilewater, followed by a lysing solution containing sodium dodecyl sulfate.The final macrophage lysate suspension is serially diluted and aliquots(0.1 mL) are plated in duplicate on Middlebrook 7H10 agar. Results canbe reported as mean numbers of colony-forming units per milliliter ofmacrophage lysate, with each assay performed in triplicate. The MIC isthe lowest concentration of drug that results in 99.9% killing.

Methodology for Testing of Legionella pneumophila

MICs are performed according to NCCLS guidelines in 96-well microtitertrays (National Committee for Clinical Laboratory Standards 1990). Ahuman monocyte cell line HL-60 (1.5×10⁶ cells/well) is infected with1.5×10⁷ colony-forming units of L. pneumophilia; after 6 h, theextracellular bacteria are removed by 4 washes, and compound added atvarying concentrations. After 48 h, cells are removed with trypsin-EDTAand cell-associated bacteria counted from duplicate wells by hypotoniclysis of the cells with sterile distilled water, followed by serialdilution and plate counts on buffered yeast extract agar containing 0.1%α-keto glutarate (Stout, J. E. et al., Diagnostic Microbiology andInfectious Disease, 1998, 30:37-43). The MIC is the lowest concentrationof drug that results in 99.9% killing.

Methodology for Testing of Toxoplasma gondii

Human foreskin fibroblast (HFF) cells (ATCC HS68) are grown inDulbecco's modified Eagle's medium (Gibco BRL, Grand Island, N.Y.)containing 100 U of penicillin, 1 μg of streptomycin per ml, and 10%heat-inactivated T. gondii antibody-negative fetal bovine serum. Invitro activity is defined as the capacity of a compound to inhibitintracellular replication of T. gondii and is determined by the[³H]uracil incorporation technique (Khan, et al., Antimicrob. AgentsChemother, 1996, 40:1855-1859). Briefly, the protocol consists ofplating HFF cells at 10⁴ cells/well in 96-well flat-bottom tissueculture microtiter plates, followed by incubation at 37° C. in a 5% CO₂incubator. After confluence, the monolayers are infected withtachyzoites at a ratio of three tachyzoites/cell. Four hours later, themonolayers are washed, compounds are added at varying concentrations,and the cultures incubated for 48 h. Four hours prior to harvesting ofthe cells, [³H]uracil (1 μCi/well) is added and its level ofincorporation determined. The cells are collected with a cell harvester,and the radioactivity counted with a scintillation counter. Compoundsare compared by their IC₅₀ values, i.e., the concentration that inhibits50% of [³H]uracil incorporation uptake and incorporation.

Methods for testing of activity against Chlamydia pneumoniae aredescribed in the Examples below.

Glycogen Phosphorylase from Mammalian Sources:

The three different purified glycogen phosphorylase (GP) isoenzymes froma human source, wherein glycogen phosphorylase is in the activated “a”state (referred to as glycogen phosphorylase a, or the abbreviationGPa), and referred to here as human liver glycogen phosphorylase a(HLGPa), human muscle glycogen phosphorylase a (HMGPa), and human brainglycogen phosphorylase a (HBGPa), can be obtained by the followingprocedures.

Expression and Fermentation

The HLGP, and HMGP cDNAs are expressed from plasmid pKK233-2 (PharmaciaBiotech. Inc., Piscataway, N.J.) in E. coli strain XL-1 Blue (StratageneCloning Systems, Lajolla, Calif.). The strain is inoculated into LBmedium (consisting of 10 g tryptone, 5 g yeast extract, 5 g NaCl, and 1ml 1N NaOH per liter) plus 100 mg/L ampicillin, 100 mg/L pyridoxine and600 mg/L MnCl₂ and grown at 37° C. to a cell density of OD₅₅₀=1.0. Atthis point, the cells are induced with 1 mMisopropyl-1-thio-β-D-galactoside (IPTG). Three hours after induction thecells are harvested by centrifugation and cell pellets are frozen at−70° C. until needed for purification.

The HBGP cDNA can be expressed by several methodologies, for example, bythe method described by Crerar, et al. (J. Biol. Chem. 270:13748-13756).The method described by Crerar, et al. for the expression of HBGP is asfollows: the HBGP cDNA can be expressed from plasmid pTACTAC in E. Colistrain 25A6. The strain is inoculated into LB medium (consisting of 10 gtryptone, 5 g yeast extract, 5 g NaCl, and 1 ml 1 N NaOH per liter) plus50 mg/L ampicillin and grown overnight, then resuspended in fresh LBmedium plus 50 mg/L ampicillin, and reinoculated into a 40× volume ofLB/amp media containing 250 μM isopropyl-1-thio-β-D-galactoside (IPTG),0.5 mM pyridoxine and 3 mM mg/L and grown at 22° C. for 48-50 hours. Thecells can then be harvested by centrifugation and cell pellets arefrozen at −70° C. until needed for purification.

The HLGP cDNA is expressed from plasmid pBlueBac III (Invitrogen Corp.,San Diego, Calif.) which is cotransfected with BaculoGold Linear ViralDNA (Pharmingen, San Diego, Calif.) into Sf9 cells. Recombinant virus issubsequently plaque-purified. For production of protein, Sf9 cells grownin serum-free medium are infected at an moi of 0.5 and at a cell densityof 2×10⁶ cells/ml. After growth for 72 hours at 27° C., cells arecentrifuged, and the cell pellets frozen at −70° C. until needed forpurification.

Purification of Mammalian Glycogen Phosphorylase Expressed in E. coli

The E. coli cells in pellets described above are resuspended in 25 mMβ-glycerophosphate (pH 7.0) with 0.2 mM DTT, 1 mM MgCl₂, plus thefollowing protease inhibitors:

0.7 μg/mL Pepstatin A 0.5 μg/mL Leupeptin 0.2 mM phenylmethylsulfonylfluoride (PMSF), and 0.5 mM EDTA,

lysed by pretreatment with 200 μg/mL lysozyme and 3 pg/mL DNAasefollowed by sonication in 250 mL batches for 5×1.5 minutes on ice usinga Branson Model 450 ultrasonic cell disrupter (Branson Sonic Power Co.,Danbury Conn). The E. coli cell lysates are then cleared bycentrifugation at 35,000×g for one hour followed by filtration through0.45 micron filters GP in the soluble fraction of the lysates (estimatedto be less than 1% of the total protein) is purified by monitoring theenzyme activity (as described in GPa Activity Assay section, below) froma series of chromatographic steps detailed below.

Immobilized Metal Affinity Chromatography (IMAC)

This step is based on the method of Luong et al. (Luong et al. Journalof Chromatography (1992) 584, 77-84). 500 mL of the filtered solublefraction of cell lysates (prepared from approximately 160-250 g oforiginal cell pellet) are loaded onto a 130 mL column of IMACChelating-Sepharose (Pharmacia LKB Biotechnology, Piscataway, N.J.)which has been charged with 50 mM CuCl₂ and 25 mM β-glycerophosphate,250 mM NaCl and 1 mM imidazole at pH 7 equilibration buffer. The columnis washed with equilibration buffer until the A280 returns to baseline.The sample is then eluted from the column with the same buffercontaining 100 mM imidazole to remove the bound GP and other boundproteins. Fractions containing the GP activity are pooled (approximately600 mL), and ethylenediaminetetraacetic acid (EDTA), DL-dithiothreitol(DTT), phenylmethylsulfonyl fluoride (PMSF), leupeptin and pepstatin Aare added to obtain 0.3 mM, 0.2 mM, 0.2 mM, 0.5 μg/mL and 0.7 μg/mLconcentrations respectively. The pooled GP is desalted over a SephadexG-25 column (Sigma Chemical Co., St. Louis, Mo.) equilibrated with 25 mMTris-HCI (pH 7.3), 3 mM DTT buffer (Buffer A) to remove imidazole and isstored on ice until the second chromatographic step.

5′- AMP-Sepharose Chromatography

The desalted pooled GP sample (approximately 600 mL) is next mixed with70 mL of 5′-AMP Sepharose (Pharmacia LKB Biotechnology, Piscataway,N.J.) which has been equilibrated with Buffer A (see above). The mixtureis gently agitated for one hour at 22° C. then packed into a column andwashed with Buffer A until the A₂₈₀ returns to baseline. GP and otherproteins are eluted from the column with 25 mM Tris-HCl, 0.2 mM DTT and10 mM adenosine 5′-monophosphate (AMP) at pH 7.3 (Buffer B).GP-containing fractions are pooled following identification bydetermining enzyme (described below) activity and visualizing the M_(r)approximately 97 kdal GP protein band by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) followed by silverstaining (2D-silver Stain II “Daiichi Kit”, Daiichi Pure Chemicals Co.,LTD., Tokyo, Japan) and then pooled. The pooled GP is dialyzed into 25mM β-glycerophosphate, 0.2 mM DTT, 0.3 mM EDTA, 200 mM NaCl, pH 7.0buffer (Buffer C) and stored on ice until use.

Prior to use of the GP enzyme, the enzyme is converted from the inactiveform as expressed in E. coli strain XL-1 Blue (designated GPb)(Stratagene Cloning Systems, La Jolla, Calif.), to the active form(designated GPa) by the procedure described in the section Activation ofGP below.

Purification of Glycogen Phosphorylase Expressed in Sf9 Cells

The Sf9 cells in pellets described above are resuspended in 25 mMβ-glycerophosphate (pH 7.0) with 0.2 mM DTT, 1 mM MgCl2, plus thefollowing protease inhibitors:

0.7 μg/mL Pepstatin A 0.5 μg/mL Leupeptin 0.2 mM phenylmethylsulfonylfluoride (PMSF), and 0.5 mM EDTA,

lysed by pretreatment with 3 μg/mL DNAase followed by sonication inbatches for 3×1 minutes on ice using a Branson Model 450 ultrasonic celldisrupter (Branson Sonic Power Co., Danbury Conn.). The Sf9 cell lysatesare then cleared by centrifugation at 35,000×g for one hour followed byfiltration through 0.45 micron filters. GP in the soluble fraction ofthe lysates (estimated to be 1.5% of the total protein) is purified bymonitoring the enzyme activity (as described in GPa Activity Assaysection, below) from a series of chromatographic steps detailed below.

Immobilized Metal Affinity Chromatography (IMAC)

Immobilized Metal Affinity Chromatography is performed as described inthe section above. The pooled, desalted GP is then stored on ice untilfurther processed.

Activation of GP

Before further chromatography, the fraction of inactive enzyme asexpressed in Sf9 cells (designated GPb) is converted to the active form(designated GPa) by the following procedure described in Activation ofGP below.

Anion Exchange Chromatography

Following activation of the IMAC purified GPb to GPa by reaction withthe immobilized phosphorylase kinase, the pooled GPa fractions aredialyzed against 25 mM Tris-HCl, pH 7.5, containing 0.5 mM DTT, 0.2 mMEDTA, 1.0 mM phenylmethylsulfonyl fluoride (PMSF), 1.0 pg/mL leupeptinand 1.0 pg/mL pepstatin A. The sample is then loaded onto a MonoQ AnionExchange Chromatography column (Pharmacia Biotech. Inc., Piscataway,N.J.). The column is washed with equilibration buffer until the A₂₈₀returns to baseline. The sample is then eluted from the column with alinear gradient of 0-0.25 M NaCl to remove the bound GP and other boundproteins. GP-containing fractions elute between 0.1-0.2 M NaCl range, asdetected by monitoring the eluant for peak protein absorbance at A₂₈₀.The GP protein is then identified by visualizing the M_(r) approximately97 kdal GP protein band by sodium dodecyl sulfate polyacrylamide gelelectrophoresis (SDS-PAGE) followed by silver staining (2D-silver StainII “Daiichi Kit”, Daiichi Pure Chemicals Co., LTD., Tokyo, Japan) andthen pooled. The pooled GP is dialyzed into 25 mM BES, 1.0 mM DTT, 0.5mM EDTA, 5 mM NaCl, pH 6.8 buffer and stored on ice until use.

Determination of GP Enzyme Activity

Activation of GP: Conversion of GPb to GPa

Prior to the determination of GP enzyme activity, the enzyme isconverted from the inactive form as expressed in E. coli strain XL-1Blue (designated GPb) (Stratagene Cloning Systems, La Jolla, Calif.), tothe active form (designated GPa) by phosphorylation of GP usingphosphorylase kinase as follows. The fraction of inactive enzyme asexpressed in Sf9 cells (designated GPb) is also converted to the activeform (designated GPa) by the follow procedure.

GP reaction with Immobilized Phosphorylase Kinase

Phosphorylase kinase (Sigma Chemical Company, St. Louis, Mo.) isimmobilized on Affi-Gel 10 (BioRad Corp., Melville, N.Y.) as per themanufacturer's instructions. In brief, the phosphorylase kinase enzyme(10 mg) is incubated with washed Affi-Gel beads (1 mL) in 2.5 mL of 100mM HEPES and 80 mM CaCl₂ at pH 7.4 for 4 hours at 4° C. The Affi-Gelbeads are then washed once with the same buffer prior to blocking with50 mM HEPES and 1 M glycine methyl ester at pH 8.0 for one hour at roomtemperature. Blocking buffer is removed and replaced with 50 mM HEPES(pH 7.4), 1 mM β-mercaptoethanol and 0.2% NaN₃ for storage. Prior to useto convert GPb to GPa, the Affi-Gel immobilized phosphorylase kinasebeads are equilibrated by washing in the buffer used to perform thekinase reaction, consisting of 25 mM β-glycerophosphate, 0.3 mM DTT, and0 3mM EDTA at pH 7.8 (kinase assay buffer).

The partially purified, inactive GPb obtained from 5′-AMP-Sepharosechromatography above (from E. coli) or the mixture of GPa and GPbobtained from IMAC above (from Sf9 cells) is diluted 1:10 with thekinase assay buffer then mixed with the aforementioned phosphorylasekinase enzyme immobilized on the Affi-Gel beads. NaATP is added to 5 mMand MgCl₂ to 6 mM. The resulting mixture is mixed gently at 25° C. for30 to 60 minutes. The sample is removed from the beads and the percentactivation of GPb by conversion to GPa is estimated by determining GPenzyme activity in the presence and absence of 3.3 mM AMP. The percentof total GP enzyme activity due to GPa enzyme activity (AMP-independent)is then calculated as follows:

((HLGP activity−AMP)/(HLGP activity+AMP)) 100

Alternately, the conversion of GPb to GPa can be monitored byisoelectric focusing based on the shift in electrophoretic mobility thatis noted following conversion of GPb to GPa GP samples are analyzed byisoelectric focusing (IEF) utilizing the Pharmacia PfastGel System(Pharmacia Biotech. Inc., Piscataway, N.J.) using precast gels (pl range4-6.5) and the manufacturer's recommended method. The resolved GPa andGPb bands are then visualized on the gels by silver staining (2D-silverStain II “Daiichi Kit”, Daiichi Pure Chemicals Co., LTD., Tokyo, Japan).Identification of GPa and GPb is made by comparison to E. coli derivedGPa and GPb standards that are run in parallel on the same gels as theexperimental samples.

GPa Activity Assay

The effect of the compounds of Formula I or IA on the activity of theactivated form of glycogen phosphorylase (GPa) can be determined by oneof two methods; glycogen phosphorylase a activity is measured in theforward direction by monitoring the production of glucose-1-phosphatefrom glycogen or by following the reverse reaction, measuring glycogensynthesis from glucose-1-phosphate by the release of inorganicphosphate. All reactions are run in triplicate in 96-well microtiterplates and the change in absorbance due to formation of the reactionproduct is measured at the wavelength specified below in a MCC/340 MKIIElisa Reader (Lab Systems, Finland), connected to a Titertech MicroplateStacker (ICN Biomedical Co, Huntsville, Ala.).

To measure the GPa enzyme activity in the forward direction, theproduction of glucose-1-phosphate from glycogen is monitored by themultienzyme coupled general method of Pesce et al. (Pesce, M. A.,Bodourian, S. H., Harris, R. C. and Nicholson, J. F. (1977) ClinicalChemistry 23, 1711-1717) modified as follows: 1 to 100 pg GPa, 10 unitsphosphoglucomutase and 15 units glucose-6-phosphate dehydrogenase(Boehringer Mannheim Biochemicals, Indianapolis, Ind.) are diluted to 1mL in Buffer A (described hereinafter). Buffer A is at pH 7.2 andcontains 50 mM HEPES, 100 mM KCl, 2.5 mM ethyleneglycoltetraacetic acid(EGTA), 2.5 mM MgCl₂, 3.5 mM KH₂PO₄ and 0.5 mM dithiothreitol. 20 μl ofthis stock is added to 80 μl of Buffer A containing 0.47 mg/mL glycogen,9.4 mM glucose, 0.63 mM of the oxidized form of nicotinamide adeninedinucleotide phosphate (NADP+). The compounds to be tested are added as5 μL of solution in 14% dimethylsulfoxide (DMSO) prior to the additionof the enzymes. The basal rate of GPa enzyme activity in the absence ofinhibitors is determined by adding 5 μL of 14% DMSO and afully-inhibited rate of GPa enzyme activity is obtained by adding 20 μLof 50 mM of the positive control test substance, caffeine. The reactionis followed at room temperature by measuring the conversion of oxidizedNADP+ to reduced NADPH at 340 nm.

To measure the GPa enzyme activity in the reverse direction, theconversion of glucose-1-phosphate into glycogen plus inorganic phosphateis measured by the general method described by Engers et al. (Engers, H.D., Shechosky, S. and Madsen, N. B. (1970) Can. J Biochem. 48, 746-754)modified as follows: 1 to 100 ug GPa is diluted to 1 mL in Buffer B(described hereinafter). Buffer B is at pH 7.2 and contains 50 mM HEPES,100 mM KCl, 2.5 mM EGTA, 2 5 mM MgCl₂ and 0.5 mM dithiothreitol. 20 μLof this stock is added to 80 μL of Buffer B with 1.25 mg/mL glycogen,9.4 mM glucose, and 0.63 mM glucose-1-phosphate. The compounds to betested are added as 5 μL of solution in 14% DMSO prior to the additionof the enzyme. The basal rate of GPa enzyme activity in the absence ofadded inhibitors is determined by adding 5 μL of 14% DMSO and afully-inhibited rate of GPa enzyme activity is obtained by adding 20 μLof 50 mM caffeine. This mixture is incubated at room temperature for 1hour and the inorganic phosphate released from the glucose-1-phosphateis measured by the general method of Lanzetta et al. (Lanzetta, P. A.,Alvarez, L. J., Reinach, P. S. and Candia, O. A. (1979) Anal. Biochem.100, 95-97) modified as follows: 150 μL of 10 mg/mL ammonium molybdate,0.38 mg/mL malachite green in 1 N HCl is added to 100 μL of the enzymemix. After a 20 minute incubation at room temperature, the absorbance ismeasured at 620 nm.

The above assays can also be used to assess activity of glycogenphosphorylase derived from various pathogenic sources. Adaptation of theassays as required is easily accomplished.

The above assays carried out with a range of concentrations of testcompound allows the determination of an IC₅₀ value (concentration oftest compound required for 50% inhibition) for the in vitro inhibitionof GPa enzyme activity by that test compound.

The inhibiting effect of compounds employed in the invention on thehuman liver and human muscle glycogen phosphorylase a isoforms is shownin Table 1 below.

TABLE 1* HLGPa HMGPa Compound Name IC₅₀ nM IC₅₀ nM5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-(2R)-  54  96hydroxy-3-((3S)-hydroxy-pyrrolidin-1-yl)-3-oxopropyl]-amide5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl(2R)-  73  90hydroxy-3-((3S,4S)-dihydroxy-pyrrolidin-1-yl)-3-oxopropyl]-amide5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-3-((3- 236 706 hydroxyazetidin-1-yl)-(2R)-hydroxy-3-oxopropyl]-amide5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-3-(cis-  59 3853,4-dihydroxy-pyrrolidin-1-yl)-(2R)-hydroxy-3-oxopropyl]-amide5-chloro-1H-indole-2-carboxylic acid [1-benzyl-2-(3-  45  85hydroxypyrrolidin-1-yl)-2-oxo-ethyl]-amide5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2-(cis-  30  973,4-dihydroxypyrrolidin-1-yl)-2-oxo-ethyl]-amide5-chloro-1H-indole-2-carboxylic acid [(1S)-(4- 142  83fluorobenzyl-2-(4-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide5-chloro-1H-indole-2-carboxylic acid (2-oxo-2-thiazolidin- 307 4333-yl-ethyl)-amide 5-chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-(3-  65 121 hydroxy-azetidin-1-yl)-2-oxo-ethyl]-amide5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2-(3-  65  84hydroxyimino-azetidin-1-yl)-2-oxo-ethyl]-amide5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2- 137  71((3S,4S)-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide *data are forHLGPa and HMGPa enzyme activity (IC₅₀) as determined by the reversedirection assay.

Generally, the glycogen phosphorylase inhibitors are administeredorally, but parenteral administration (e.g., intravenous, intramuscular,subcutaneous or intramedullary) may be utilized, for example, where oraladministration is inappropriate or where the patient is unable to ingestthe drug. For certain tissues such as the eye, topical administrationmay also be suitable.

The glycogen phosphorylase inhibitors may be administered alone or incombination with pharmaceutically acceptable carriers, in either singleor multiple doses. Suitable pharmaceutical carriers include inert soliddiluents or fillers, sterile aqueous solutions, oils (e.g., peanut oil,sesame oil) and various organic solvents. The pharmaceuticalcompositions formed by combining the active compounds andpharmaceutically acceptable carriers can then be readily administered ina variety of dosage forms such as tablets, powders, lozenges, emulsions,oil soft gels, syrups, injectable solutions, spray-dried formulations,transdermal or transmucosal patches, inhalable formulations and thelike. These pharmaceutical compositions can, if desired, containadditional ingredients such as flavorings, binders, excipients and thelike. Thus, for purposes of oral administration, tablets containingvarious excipients such as sodium citrate, calcium carbonate and calciumphosphate may be employed along with various disintegrants such asstarch, methylcellulose, alginic acid and certain complex silicates,together with binding agents such as polyvinylpyrrolidone, sucrose,gelatin and acacia. Additionally, lubricating agents such as magnesiumstearate, sodium lauryl sulfate and talc are often useful for tablettingpurposes. Solid compositions of a similar type may also be employed asfillers in soft and hard filled gelatin capsules. Preferred materialsfor this include lactose or milk sugar and high molecular weightpolyethylene glycols. When aqueous suspensions or elixirs are desiredfor oral administration, the essential active ingredient therein may becombined with various sweetening or flavoring agents, coloring matter ordyes and, if desired, emulsifying or suspending agents, together withdiluents such as water, ethanol, propylene glycol, glycerin andcombinations thereof.

For parenteral administration, solutions containing an active compoundor a pharmaceutically acceptable salt thereof in sesame or peanut oil,aqueous propylene glycol, or in sterile aqueous solution may beemployed. Such aqueous solutions should be suitably buffered ifnecessary and the liquid diluent first rendered isotonic with sufficientsaline or glucose. These particular aqueous solutions are especiallysuitable for intravenous, intramuscular, subcutaneous andintraperitoneal administration. The sterile aqueous media employed areall readily available by standard techniques known to those skilled inthe art.

Methods of preparing various pharmaceutical compositions with a certainamount of active ingredient are known, or will be apparent in light ofthis disclosure, to those skilled in this art. For examples of how toprepare such compositions see Remington's Pharmaceutical Sciences, MackPublishing Company, Easton, Pa., 19th Edition (1995).

Pharmaceutical compositions administered according to the inventiongenerally contain 0.01%-95% of glycogen phosphorylase inhibitor,preferably 1%-70%. In any event, the composition or formulation to beadministered contains a quantity of a glycogen phosphorylase inhibitorin an amount effective to treat infection. Typically, an effectivedosage for the glycogen phosphorylase inhibitor is in the range of about0.005 to 50 mg/kg/day, preferably 0.01 to 25 mg/kg/day and mostpreferably 0.1 to 15 mg/kg/day.

The present invention encompasses treating or preventing infection byadministering a compound of formula I in combination with a secondcompound for treating the infection. The second compound for treatingthe infection can be, for example, an antibiotic such as anaminoglycoside, penicillin, beta-lactamase inhibitor, anti-tuberculosisagent, cephalosporin, carbapenem, quinolone, macrolide, ketolide,oxazolidinone (i.e., linezolid), streptogramins, anti-staphylococcalagent, lincosamine, sulfonamide, or other type of antibiotic. Examplesof such antibiotics include but are not limited to amoxicillin,ampicillin, polycillin, azithromycin, azlocillin, aztrenam,bacampicillin, bacitracin, benethamine, benza-thine, bicillin,benzylpenicillin, capreomycin, carbenicillin, cefadroxil, cefamandole,cefazolin, cefixime, cefizoxime, ceflacor, cefmetazole, cefoperazone,cefotaxime, cefotetan, cefoxitin, ceftazidime, ceftriaxone, cefuroxime,cephalexin, cephalothin, cephapirin, cephradine, chlorampenicol,chlortetracycline, cilastatin, ciprofloxacin, clarithromycin, clavulanicacid, clindamycin, colistin, cycloserine, dalfopristin, demeclocycline,dicloxacillin, doxycycline, erythrocin, erythromycin, ethambutol,ethionamide, fosfomycin, gentamicin, imipenem, isoniazid, kanamycin,lincomycin, linezolid, meropenem, methacycline, methenamine,mandelamine, methicillin, metronidazole, mezlocillin, minocycline,mupirocin, nafcillin, nalidixic acid, neomycin, netilmicin,nitrofurantoin, norfloxacin, novobiocin, ofloxacin, oxacillin, oxolinicacid, oxytetracycline, quinipristin, paromomycin, pefloxacin,phenoxymethylpenicillin, piperacillin, polymyxin b, procaine penicillin,pyrazinamide, r-aminosalicyclic acid, rifampin, spectinomycin,streptomycin, sulfacytine, sulfisoxazole, sullbacatam, Synercid,telithromycin, sulfadiazine, sulfamethizole, sulfamethoxazole,sulfapyridine, sulfasalazine, sulfisoxazole, sullbacatam, tetracycline,thienamycin, ticarcillin, ticarcillin, tobramycin, trimethoprim,trisulfapyrimidines, trovafloxicin, and vancomycin. Administration ofthese compounds can be carried out using dosages and formulations thatare well-known.

The invention is illustrated by the following Example, which is providedto exemplify the invention, and not to be interpreted as narrowing itsscope.

EXAMPLE

TABLE 2 Use of GP Inhibitors to treat Chlamydia pneumoniae infectionProtocol 1 Protocol 2 Compound MIC (μg/ml) MBC (μg/ml) MIC (μg/ml) 112.5 25 12.5 2 25 25 50

Compounds used in this Example are shown below:

Protocol 1 was performed in 96-well flat-bottom microtiter plates. Eachwell contained 100 μL of 10⁶ HEp-G2 cells/ml in standard medium that hadbeen incubated for 24 h at 37° C. in 5% CO₂. Compound stocks wereprepared in dimethylsulfoxide, diluted in two-fold serial dilutions, andadded to each well as 10 μL aliquots. Each compound concentration wasassayed in triplicate. Elementary bodies (chlamydial stock) were dilutedto contain 2×10⁴inclusion-forming units (ifu) per milliliter and 90 μLwas added to each well. Infection was allowed to proceed for 72 h at 37°C. in 5% CO₂, after which the cells were fixed and stained withgenus-reactive anti-LPS antibody. The use of a fluorescein(FITC)-conjugated secondary antibody allowed the number ofinclusion-containing cells to be identified via inverted fluorescentmicroscopy. The minimum inhibitory concentration (MIC) was defined asthe lowest concentration of compound that inhibited the formation ofinclusions. The minimal bactericidal concentration (MBC) was defined asthe lowest concentration of compound that prevented the formation ofinclusions after the compound had been removed by the addition of freshmedium and the cultures had been incubated for a further 48 h.Procedures for growth and preparation of C. pneumoniae and HEp-G2 cellsare described in Kalayoglu, M. V., et al., J. Infect. Dis., 1999,180:780-790. Methodology for detecting inclusions using an directfluorescent antibody technique is found in Byrne, G. I., et al. J.Infect. Dis., 1993, 168:415-420.

Protocol 2 was nearly identical to protocol 1 except that compound wasadded 15 h after challenge of HEp-G2 cells with C. pneumoniae. Thisprotocol helps distinguish compounds that interfere with latter stagesof C. pneumoniae growth and replication.

As shown in Table 2, both compound I and compound 2 exhibited activityagainst growth of C. pneumoniae in HEp-2 cells. Compound 1 was superiorto compound 2 in interfering with the latter stage of pneumoniae growthand replication. Also of note was that the MICs of compound I andcompound 2 did not differ in a protocol that treats HEp-G2 cells withcycloheximide (1 μg/mL).

What is claimed is:
 1. A method of treating a Chiamydia pneumoniae or aChlamydia trachomatis infection in a mammal comprising administering aneffective amount of a glycogen phosphorylase inhibitor of Formula I orFormula IA

and the pharmaceutically acceptable salts and prodrugs thereof; wherein:the dotted line (---) is an optional bond; A is —C(H)═,—C((C₁-C₄)alkyl)═ or —C(halo)═ when the dotted line (---) is a bond, orA is methylene or —CH((C₁-C₄)alkyl)- when the dotted line (---) is not abond; R₁, R₈ or R₉ are each independently H, halo, 4-, 6- or 7-nitro,cyano, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, fluoromethyl, difluoromethyl ortrifluoromethyl; R₂ is H; R₃ is H or (C₁-C₅)alkyl; R₄ is H, methyl,ethyl, n-propyl, hydroxy(C₁-C₃)alkyl, (C₁-C₃)alkoxy(C₁-C₃)alkyl,phenyl(C₁-C₄)alkyl, phenylhydroxy(C₁-C₄)alkyl,phenyl(C₁-C₄)alkoxy(C₁-C₄)alkyl, thien-2- or -3-yl(C₁-C₄)alkyl or fur-2-or - 3-yl(C₁-C₄)alkyl wherein said R₄ rings are mono-, di- ortri-substituted independently on carbon with H, halo, (C₁-C₄)alkyl,(C₁-C₄)alkoxy, trifluoromethyl, hydroxy, amino or cyano; or R₄ ispyrid-2-, -3- or -4-yl(C₁-C₄)alkyl, thiazol-2-, -4- or-5-yl(C₁-C₄)alkyl, imidazol -1-, -2-, -4- or - 5-yl(C₁-C₄)alkyl,pyrrol-2- or -3-yl(C₁-C₄)alkyl, oxazol-2-, - 4- or -5-yl-(C₁-C₄)alkyl,pyrazol-3-, -4- or -5- yl(C₁-C₄)alkyl, isoxazol-3-, -4- or-5-yl(C₁-C₄)alkyl, isothiazol-3-, -4- or -5-yl(C₁-C₄)alkyl, pyridazin-3-or -4-yl-(C₁-C₄)alkyl, pyrimidin-2-, -4-, -5- or -6- yl(C₁-C₄)alkyl,pyrazin-2- or -3-yl(C₁-C₄)alkyl or 1,3,5-triazin-2-yl(C₁-C₄)alkyl,wherein said preceding R₄ heterocycles are optionally mono- ordi-substituted independently with halo, trifluoromethyl, (C₁-C₄)alkyl,(C₁-C₄)alkoxy, amino or hydroxy and said mono-or di- substituents arebonded to carbon; R₅ is H, hydroxy, fluoro, (C₁-C₅)alkyl, (C₁-C₅)alkoxy,(C₁-C₆)alkanoyl, amino(C₁-C₄)alkoxy, mono-N- or di-N,N-(C₁-C₄)alkylamino(C₁-C₄)alkoxy, carboxy(C₁-C₄)alkoxy,(C₁-C₅)alkoxy-carbonyl(C₁-C₄)alkoxy, benzyloxycarbonyl(C₁-C₄)alkoxy, orcarbonyloxy wherein said carbonyloxy is carbon-carbon linked withphenyl, thiazolyl, imidazolyl, 1H-indolyl, furyl, pyrrolyl, oxazolyl,pyrazolyl, isoxazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinylor 1,3,5-triazinyl and wherein said preceding R₅ rings are optionallymono-substituted with halo, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, hydroxy, aminoor trifluoromethyl and said mono-substituents are bonded to carbon; R₇is H, fluoro or (C₁-C₅)alkyl; or R₅ and R₇ can be taken together to beoxo; R₆ is C(O)R₁₀; R₁₀ is piperazin-1-yl, 4-(C₁-C₄)alkylpiperazin-l-yl, 4-formylpiperazin-1-yl, morpholino,thiomorpholino, 1-oxothiomorpholino, 1,1-dioxo-thiomorpholino,thiazolidin-3-yl, 1-oxo-thiazolidin-3-yl, 1,1-dioxo-thiazolidin-3-yl,2-(C₁-C₆)alkoxycarbonylpyrrolidin-1-yl, oxazolidin-3-yl or 2(R)-hydroxymethylpyrrolidin-1-yl; or R₁₀ is 3- and/or 4-mono-ordi-substituted oxazetidin-2-yl, 2-, 4-, and/or 5- mono- ordi-substituted oxazolidin-3-yl, 2-, 4-, and/or 5- mono- ordi-substituted thiazolidin-3-yl, 2-, 4-, and/or 5- mono-or di-substituted 1-oxothiazolidin-3-yl, 2-, 4-, and/or 5- mono- or di-substituted 1,1-dioxothiazolidin-3-yl, 3- and/or 4-, mono- ordi-substituted pyrrolidin-1-yl, 3-, 4- and/or 5-, mono-, di- ortri-substituted piperidin-1-yl, 3-, 4-, and/or 5- mono-, di-, ortri-substituted piperazin-1-yl, 3-substituted azetidin-1-yl, 4- and/or5-, mono- or di-substituted 1,2-oxazinan-2-yl, 3-and/or 4-mono- ordi-substituted pyrazolidin-1-yl, 4- and/or 5-, mono- or di-substitutedisoxazolidin-2-yl, 4- and/or 5-, mono- and/or di-substitutedisothiazolidin-2-yl wherein said R₁₀ substituents are independently H,halo, (C₁-C₅)-alkyl, hydroxy, amino, mono-N- or di-N,N-(C₁-C₅)alkylamino, formyl, oxo, hydroxyimino, (C₁-C₅)alkoxy, carboxy,carbamoyl, mono-N-or di-N,N-(C₁-C₄)alkylcarbamoyl, (C₁-C₄)alkoxyimino,(C₁-C₄)alkoxymethoxy, (C₁-C₆)alkoxycarbonyl, carboxy (C₁-C₅)alkyl orhydroxy(C₁-C₅)alkyl; R₁₂ is H, methyl, ethyl, n-propyl,hydroxy(C₁-C₃)alkyl, (C₁-C₃)alkoxy(C₁-C₃)alkyl, phenyl(C₁-C₄)alkyl,phenylhydroxy(C₁-C₄)alkyl, (phenyl) ((C₁-C₄)-alkoxy) (C₁-C₄)alkyl,thien-2- or -3-yl(C₁-C₄)alkyl or fur-2- or -3-yl(C₁-C₄)alkyl whereinsaid R₄ rings are mono-, di- or tri-substituted independently on carbonwith H, halo, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, trifluoromethyl, hydroxy,amino, cyano or 4,5-dihydro-1H-imidazol-2-yl; or R₁₂ is pyrid-2-, -3- or-4-yl(C₁-C₄)alkyl, thiazol-2-, -4- or -5-yl(C₁-C₄)alkyl, imidazol-2-,-4- or -5-yl(C₁-C₄)alkyl, pyrrol-2- or -3-yl(C₁-C₄)alkyl, oxazol-2-, 4-or -5-yl(C₁-C₄)alkyl, pyrazol-3-, -4- or -5-yl(C₁-C₄)alkyl, isoxazol-3-,-4- or -5-yl(C₁-C₄)alkyl, isothiazol-3-, -4- or -5-yl(C₁-C₄)alkyl,pyridazin-3-or -4-yl(C₁-C₄)alkyl, pyrimidin-2-, -4-, -5- or-6-yl(C₁-C₄)alkyl, pyrazin-2- or -3-yl(C₁-C₄)alkyl,1,3,5-triazin-2-yl(C₁-C₄)alkyl or indol-2-(C₁-C₄)alkyl, wherein saidpreceding R₁₂ heterocycles are optionally mono- or di-substitutedindependently with halo, trifluoromethyl, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,amino, hydroxy or cyano and said substituents are bonded to carbon; orR₁₂ is R₁₁-carbonyloxymethyl, wherein said R₁₁ is phenyl, thiazolyl,imidazolyl, 1H-indolyl, furyl, pyrrolyl, oxazolyl, pyrazolyl,isoxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinylor 1,3,5-triazinyl and wherein said preceding R₁₁ rings are optionallymono- or di-substituted independently with halo, amino, hydroxy,(C₁-C₄)alkyl, (C₁-C₄)alkoxy or trifluoromethyl and said mono- ordi-substituents are bonded to carbon; R₁₃ is H; R₁₄ C(O)R₁₅; R₁₅ ismorpholino, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, thiazolidin-3-yl, 1-oxothiazolidin-3-yl,1,1-dioxothiazolidin-3-yl, pyrrolidin-1-yl, piperidin-1-yl,piperazin-1-yl, piperazin-4-yl, azetidin-1-yl, 1,2-oxazinan-2-yl,pyrazolidin-1-yl, isoxazolidin-2-yl, isothiazolidin-2-yl,1,2-oxazetidin-2-yl, oxazolidin-3-yl, 3,4-dihydroisoquinolin-2-yl,1,3-dihydroisoindol-2-yl, 3,4-dihydro-2H-quinol-1-yl,2,3-dihydro-benzo[1,4]oxazin-4-yl, 2,3-dihydro-benzo[1,4]-thiazine-4-yl,3,4-dihydro-2H-quinoxalin-1-yl, 3,4-dihydro-benzo[c][1,2]oxazin-1-yl,1,4-dihydro-benzo[d][1,2]oxazin-3-yl, 3,4-dihydro-benzo[e][1,2]-oxazin-2-yl, 3H-benzo[d]isoxazol-2-yl, 3H-benzo[c]isoxazol-1-yl or azepan-1-yl,wherein said R₁₅ rings are optionally mono-, di- or tri-substitutedindependently with halo, (C₁-C₅)alkyl, (C₁-C₅)alkoxy, hydroxy, amino,mono-N- or di-N,N-(C₁-C₅)alkylamino, formyl, carboxy, carbamoyl, mono-N-or di-N,N- (C₁-C₅)alkylcarbamoyl, (C₁-C₆)alkoxy(C₁-C₃)alkoxy,(C₁-C₅)alkoxycarbonyl, benzyloxycarbonyl, (C₁-C₅)alkoxycarbonyl(C₁-C₅)alkyl, (C₁-C₄)alkoxycarbonylamino, carboxy(C₁-C₅)alkyl,carbamoyl(C₁-C₅)alkyl, mono-N- ordi-N,N-(C₁-C₅)alkylcarbamoyl(C₁-C₅)alkyl, hydroxy(C₁-C₅)alkyl,(C₁-C₄)alkoxy(C₁-C₄)alkyl, amino(C₁-C₄)alkyl, mono-N- or di-N,N-(C₁-C₄)alkylamino(C₁-C₄)alkyl, oxo, hydroxyimino or (C₁-C₆)alkoxyiminoand wherein no more than two substituents are selected from oxo,hydroxyimino or (C₁-C₆)alkoxyimino and oxo, hydroxyimino or(C₁-C₆)alkoxyimino are on nonaromatic carbon; and wherein said R₁₅ ringsare optionally additionally mono- or di-substituted independently with(C₁-C₅)alkyl or halo.
 2. A method according to claim 1 wherein: R₁ is5-H, 5-halo, 5-methyl or 5-cyano; R₈ and R₉ are each independently H orhalo; A is —C(H)═; R₂ and R₃ are H; R₄ is phenyl(C₁-C₂)alkyl whereinsaid phenyl groups are mono-, di- or tri-substituted independently withH or halo or mono- or di- substituted independently with H, halo,(C₁-C₄)alkyl, (C₁-C₄)alkoxy, trifluoromethyl, hydroxy, amino or cyano;or R₄ is thien-2- or -3-yl(C₁-C₂)alkyl, pyrid-2-, -3- or-4-yl(C₁-C₂)alkyl, thiazol-2-, -4- or -5-yl(C₁-C₂)alkyl, imidazol -1-,-2-, -4- or -5-yl(C₁-C₂)alkyl, fur-2- or -3-yl(C₁-C₂)alkyl, pyrrol-2- or-3-yl(C₁-C₂)alkyl, oxazol-2-, -4- or -5-yl-(C₁-C₂)alkyl, pyrazol-3-, -4-or -5-yl(C₁-C₂)alkyl, isoxazol-3-, -4- or -5-yl(C₁-C₂)alkyl wherein saidpreceding R₄ heterocycles are optionally mono- or di-substitutedindependently with halo, trifluoromethyl, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,amino or hydroxy and said mono- or di-substituents are bonded to carbon;R₅ is hydroxy; and R₇ is H.
 3. A method according to claim 2 wherein:the carbon atom labelled a has (S) stereochemistry; the carbon atomlabelled b has (R) stereochemistry; R₄ is phenyl(C₁-C₂)alkyl,thien-2-yl-(C₁-C₂)alkyl, thien-3-yl-(C₁-C₂)alkyl, fur-2-yl-(C₁-C₂)alkylor fur-3-yl-(C₁-C₂)alkyl wherein said rings are mono- or di- substitutedindependently with H or fluoro; and R₁₀ is morpholino,4-(C₁-C₄)alkylpiperazin-1-yl, 3-substituted azetidin-1-yl, 3- and/or 4-,mono- or di-substituted pyrrolidin-1-yl, 4- and/or 5- mono- ordi-substituted isoxazolidin-2-yl, 4- and/or 5-, mono- or di-substituted1,2-oxazinan-2-yl wherein said substituents are each independently H,halo, hydroxy, amino, mono-N- or di-N,N-(C₁-C₆)alkylamino, oxo,hydroxyimino or alkoxy.
 4. A method according to claim 1 wherein thecompound is of Formula I and wherein: R₁ is H, halo, methyl or cyano; R₈and R₉ are each independently H or halo; A is —C(H)═; R₂ and R₃ are H;R₄ is phenyl(C₁-C₂)alkyl wherein said phenyl groups are mono-, di- ortri-substituted independently with H or halo or mono- or di- substitutedindependently with H, halo, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,trifluoromethyl, hydroxy, amino or cyano; or R₄ is thien-2- or-3-yl(C₁-C₂)alkyl, pyrid-2-, -3- or -4-yl(C₁-C₂)alkyl, thiazol-2-, -4-or -5-yl(C₁-C₂)alkyl, imidazol -1-, -2-, -4- or -5-yl(C₁-C₂)alkyl,fur-2- or -3-yl(C₁-C₂)alkyl, pyrrol-2- or -3-yl(C₁-C₂)alkyl, oxazol-2-,-4- or -5-yl-(C₁-C₂)alkyl, pyrazol-3-, -4- or -5-yl(C₁-C₂)alkyl,isoxazol-3-, -4- or -5-yl(C₁-C₂)alkyl wherein said preceding R₄heterocycles are optionally mono- or di-substituted independently withhalo, trifluoromethyl, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, amino or hydroxy andsaid mono- or di-substituents are bonded to carbon; R₅ is fluoro,(C₁-C₄)alkyl, (C₁-C₅)alkoxy, amino(C₁-C₄)alkoxy, mono-N- ordi-N,N-(C₁-C₄)alkylamino(C₁-C₄)alkoxy, carboxy(C₁-C₄)alkoxy,(C₁-C₅)alkoxy-carbonyl(C₁-C₄)alkoxy, benzyloxycarbonyl(C₁-C₄)alkoxy; andR₇ is H, fluoro or (C₁-C₆)alkyl.
 5. A method according to claim 1wherein said compound is selected from the group consisting of:5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-(2R)-hydroxy-3-(4-methylpiperazin-1-yl)-3-oxo-propyl]-amidehydrochloride, 5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-(2R)-hydroxy-3-(3-hydroxyazetidin-1-yl)-3-oxo-propyl]-amide,5-Chloro-1H-indole-2-carboxylic acid((1S)-benzyl-(2R)-hydroxy-3-isoxazolidin-2-yl-3-oxo-propyl)-amide,5-Chloro-1H-indole-2-carboxylic acid((1S)-benzyl-(2R)-hydroxy-3-[1,2]oxazinan-2-yl-3-oxo-propyl)-amide,5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-(2R)-hydroxy-3-((3S)-hydroxypyrrolidin-1-yl)-3-oxo-propyl]-amide,5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-3-((3S,4S)-dihydroxy-pyrrolidin-1-yl)-(2R)-hydroxy-3-oxo-propyl]-amide,5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-3-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-(2R)-hydroxy-3-oxo-propyl]-amide;and 5-Chloro-1H-indole-2-carboxylic acid((1S)-benzyl-(2R)-hydroxy-3-morpholin-4-yl-3-oxo-propyl)-amide.
 6. Amethod according to claim 1 wherein the compound is of the Formula IA,and wherein: R₁ is 5-H, 5-halo, 5-methyl, 5-cyano or 5-trifluoromethyl;R₈ and R₉ are each independently H or halo; A is —C(H)═; R₂ and R₃ areH; and R₁₂ is H, methyl, phenyl(C₁-C₂)alkyl, wherein said phenyl groupsare mono- or di-substituted independently with H, halo, (C₁-C₄)alkyl,(C₁-C₄)alkoxy, trifluoromethyl, hydroxy, amino or cyano and wherein saidR₁₂ groups are optionally additionally mono- substituted with halo; orR₁₂ is thien-2- or -3-yl(C₁-C₂)alkyl, pyrid-2-, -3- or-4-yl(C₁-C₂)alkyl, thiazol-2-, -4- or -5-yl(C₁-C₂)alkyl, imidazol-2-,-4- or -5-yl(C₁-C₂)alkyl, fur-2- or -3-yl(C₁-C₂)alkyl, pyrrol-2- or-3-yl(C₁-C₂)alkyl, oxazol-2-, - 4- or -5-yl(C₁-C₂)alkyl, pyrazol-3-, -4-or -5-yl(C₁-C₂)alkyl, isoxazol-3-, -4- or -5-yl(C₁-C₂)alkyl,isothiazol-3-, -4- or -5-yl(C₁-C₂)alkyl, pyridazin-3- or-4-yl(C₁-C₂)alkyl, pyrimidin-2-, -4-, -5- or -6-yl(C₁-C₂)alkyl,pyrazin-2- or -3-yl(C₁-C₂)alkyl or 1,3,5-triazin-2-yl(C₁-C₂)alkylwherein said preceding R₁₂ heterocycles are optionally mono- ordi-substituted independently with halo, trifluoromethyl, (C₁-C₄)alkyl,(C₁-C₄)alkoxy, amino or hydroxy and said mono- or di- substituents arebonded to carbon.
 7. A method according to claim 1 wherein said compoundis selected from the group consisting of:5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-(3-hydroxyimino-pyrrolidin-1 -yl)-2-oxo-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid [2-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide, 5-Chloro-1H-indole-2-carboxylic acid[2-((3S,4S)-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid[2-(1,1-dioxo-thiazolidin-3-yl)-2-oxo-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid(2-oxo-2-thiazolidin-3-yl-ethyl)-amide, 5-Chloro-1H-indole-2-carboxylicacid[(1S)-(4-fluoro-benzyl)-2-(4-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-((3RS)-hydroxy-piperidin-1 -yl)-2-oxo-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid[2-oxo-2-((1RS)-oxo-1-thiazolidin-3-yl)-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid[(1S)-(2-fluoro-benzyl)-2-(4-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-((3S,4S)-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-(3-hydroxy-azetidin-1-yl)-2-oxo-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid [(1S)-benzyl-2-(3-hydroxyimino-azetidin-1-yl)-2-oxo-ethyl]-amide,5-Chloro-1H-indole-2-carboxylic acid[(1S)-benzyl-2-(4-hydroxyimino-piperidin-1-yl)-2-oxo-ethyl]-amide, and5-Chloro-1H-indole-2-carboxylic acid[1-benzyl-2-(3-hydroxypyrrolidin-1-yl)-2-oxo-ethyl]amide.